| Summary: | Pork contamination often found in various processed meat. DNA-based
technique detection is one of the quick and accurate method for identifying
porcine contamination. The success of this technique is determined by the
capture of genomic DNA from the sample. Processed meat consist of a variety of
complex foodstuffs, so requiring specific methods for the DNA isolation. The aims
of this study was to determine the appropriate isolation DNA method for applied
to the meatballs, sausage, and shredded meat, and can be used as bases for
detection of porcine adulteration in processed meat. TNES, Sambrook with
modified and CTAB method were used in this study. Samples uses for the
experiment were meatballs, sausage and shredded meat with the addition of
pork. DNA concentration and purity was measured by spectrophotometer at λ
260 and λ280 nm. DNA concentration and purity data were analyzed by analysis
of variance based on one way completely randomized design (CRD), followed by
Duncan's New Multiple Range Test using SPSS version 16. Confirmation of pork
contamination was done by PCR-RFLP analysis, amplification of cytochrome b
gene using the primers L14841 and H15149. The results showed that Sambrook
with modified method gave the best results, the DNA bands generated band.
DNA concentrations and purity were obtained from three methods shown
significantly different (P<0.05). PCR amplification of the cytochrome b gene
produced 359 bp. The results of digestion produced fragments of size 131, 228
and 359 bp for samples containing pork, while containing no pork 359 bp.
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